Cervical cancer is a global health problem that continues to result in over 250,000 deaths a year, with 80% occurring in developing countries. Early detection of cervical abnormalities using the Pap smear has proven successful in reducing the number of deaths in industrialized nations. The high death toll from cervical cancer that still persists in developing countries could be substantially decreased if appropriate screening methods for cervical neoplasia were available in low resource setting. Widespread introduction of HPV vaccines in developing countries is not possible to date, and it appears as generally accepted that vaccination and screening should be applied in concert. Infection with a high-risk type Human papillomavirus (HPV) is correlated with >99% of cervical cancers. More than 90% of HPV infections, however, are cleared by the body without clinical consequence. Hence, diagnostic tests that merely detect the presence of HPV may result in unnecessary treatment of cervical neoplasia. Accumulating data strongly suggests that expression of HPV E6 and E7 oncoproteins is required for initiation and maintenance of cervical epithelial cell transformation. Thus, the E6 protein is an attractive diagnostic marker because it is correlated with the cancerous phenotype. Under a SBIR Phase 1 Grant, Arbor Vita has, in collaboration with PATH, developed a prototype of a HPV-E6 based lateral flow diagnostic test for cervical pre-cancer and cancer. The test exploits the ability of high-risk (oncogenic) HPV E6 oncoproteins, but not low-risk E6 proteins, to bind cellular PDZ domains. A recombinant PDZ domain is used for E6 capture, and detection occurs via anti E6 antibodies. The current prototype can detect HPV16-E6 and HPV18-E6 from as little as 5,000 cervical cancer cells in the background of cervical swab samples, and feasibility has been demonstrated for the remaining 5 of the top 7 most prevalent oncogenic HPV types. Arbor Vita and PATH now propose the development of the "E6 lateral flow cervical cancer test" towards a commercial prototype. Specific Aims of our proposal are: (1) to generate a multiplex E6 lateral flow cervical cancer prototype test for multiplex detection of E6 protein from HPV types 16, 18, 31, 33, 45, 52, 58;(2) to develop a commercial prototype that is aligned with medical practice of women's health care in the target markets;(3) to demonstrate pre-clinical validity of the commercial prototype;and (4) to develop the "E6 lateral flow cervical cancer test" towards manufacturing under GMP specifications. If successful, this diagnostic test will fill a critical need for a rapid, accurate and low cost diagnostic test that can be used at the point of care. PUBLIC HEALTH RELEVANCE: The high death toll from cervical cancer in developing countries could be reduced drastically if robust, simple to perform and cost effective screening methods were available. Under this application, Arbor Vita Corporation and PATH propose the development of a HPV-E6 oncoprotein based lateral flow cervical cancer rapid diagnostic test towards a commercial prototype that is ready for clinical trials and GMP manufacturing.